GC-TOFMS方法对彭水盐肤木蜂蜜指纹图谱的初步分析

采用气相色谱-飞行时间质谱(GC-TOFMS)的代谢组学方法,对彭水盐肤木蜂蜜的有机成分进行测定分析,共检测出83种有机物,主要为酸类、氨基酸类和芳香族类物质。所检测的两个样品H1和H2,H1所测得的有机成分有81种,H2有62种,H1、H2共同拥有的有机物60种。不同的有机物有N-乙酰色氨酸、L-色氨酸、焦谷氨酸、鸟氨酸、L-蛋氨酸、L-α-氨基丁酸、庚二酸、乙基甲基乙酸、延胡索酸、戊二酸、烟酸、衣康酸、甲基琥珀酸、丙二酸、2-甲基丁酸、月桂酸、柠康酸、吲哚-3-乙酰胺、羟基苯基乳酸、3-羟基苯乙酸、3-吲哚乙腈、邻羟基苯乙酸和嘌呤。     

Comparative proteomic analysis of livers from ketotic cows

The aim of this study was to examine molecular pathways activated in the liver of the ketotic cow to gain further understanding of the pathogenic effects of ketosis. We used two-dimensional-electrophoresis (2D-E) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry to compare the liver proteomic profile between ketotic and normal cows. Thirty-eight different proteins were identified between these groups by 2D-E gel. These proteins play a role in energy metabolism, carbohydrate degradation, fatty acid metabolism, amino acid metabolism, antioxidation, cell structure, nucleotide metabolism, and protein metabolism. This is the first time the majority of these proteins have been reported in cow ketosis. Possible explanations for the differential expression patterns and their relevance to the pathogenesis of ketosis in the cow are discussed. This study is the first to provide a detailed insight into protein expression patterns in the liver of the ketotic cow, and the different proteins involved in the pathogenesis of bovine ketosis.     

Effects of high ambient temperature on plasma metabolomic profiles in chicks

Exposure to high ambient temperature is detrimental to the poultry industry. To understand the influence from a metabolic perspective, we investigated the effects of exposure to high ambient temperature on plasma low‐molecular‐weight metabolite levels in chicks using gas chromatography/mass spectrometry‐based non‐targeted metabolomic analysis. Heat exposure for 4 days suppressed growth and food intake. Of the 92 metabolites identified, the levels of 29 decreased, whereas the levels of nine increased. We performed an enrichment analysis on the identified metabolites and found 35 candidates for metabolic processes affected by heat exposure. Among them, the sulfur amino acid metabolic pathway was clearly detected and the levels of the following related metabolites were decreased: cystathionine, cysteine, cystine, homocysteine and hypotaurine. Changes in the kynurenine pathway in tryptophan metabolism, which is linked to the immune system and oxidative stress, were also observed: kynurenine and quinolinic acid levels increased, whereas nicotinamide levels decreased. These results suggest the possible involvement of various metabolic processes in heat‐exposed chicks. Some of these metabolites would be important to understand the mechanism of biological responses to high ambient temperature in chicks.     

Some aspects of lipid metabolism in fatty liver and kidney syndrome in chicks.

Various aspects of lipid metabolism were examined in broiler chicks affected with fatty liver and kidney syndrome (FLKS). Plasma free fatty acid concentrations were invariably elevated. Plasma triglyceride concentrations were increased amounts of triglyceride-rich lipoproteins. Lipoprotein lipase activity in adipose tissue was considerably reduced, but in heart tissue the enzyme activity was increased. Hepatic lipogenesis was reduced. Rates of oxidation of palmitic and succinic acids by liver, heart and kidney were normal. The increased oxidation rate of palmitic acid following the addition of carnitine was also normal. These findings indicate that elevated blood lipid levels are likely to be an important factor contributing to the development of fatty deposition, particularly in extrahepatic tissues.     

混合油脂对清远麻鸡胸肌脂肪酸组成的影响及其代谢组机制研究

本试验旨在研究单一和混合油脂日粮对肉鸡生长性能和胸肌脂肪酸组成的影响，并利用代谢组学技术探究日粮油脂干预后胸肌脂肪酸组成发生变化的代谢机制，为调控肉鸡脂肪酸组成的研究提供新思路。选取1日龄慢速型清远麻母鸡480只，随机分为2组（每组6个重复，每个重复40只），对照组（CON）饲喂大豆油日粮，试验组（BOF）饲喂等量的混合油脂（大豆油∶猪油∶鱼油∶椰子油=1.0∶1.0∶0.5∶0.5）日粮，试验期127 d。结果表明，与对照组相比，混合油脂日粮对肉鸡平均日增重没有显著影响（P > 0.05），但显著降低了肉鸡的平均日采食量和料重比（P < 0.05）；显著增加了肉鸡胸肌中月桂酸（C12∶0）、肉豆蔻酸（C14∶0）、油酸（C18∶1n-9）、二十碳五烯酸（EPA，C20∶5n-3）和二十二碳六烯酸（DHA，C22∶6n-3）的比例，显著降低了亚油酸（C18∶2n-6）比例（P < 0.05）。利用液相色谱－质谱联用（LC-MS）技术，在试验鸡胸肌中共筛选到110种差异代谢物（变量权重值（VIP） > 1，P < 0.05），其中63种差异物质被注释，主要是磷脂代谢物，包括磷脂酰胆碱（PC）、磷脂酰丝氨酸（PS）和磷脂酰乙醇胺（PE）等。此外，与对照组相比，混合油脂组显著增加了胸肌中谷胱甘肽和肌肽含量（P < 0.05）。代谢通路富集分析表明，α-亚麻酸代谢、花生四烯酸代谢、亚油酸代谢和甘油磷酸代谢等脂质代谢通路发生显著改变（P < 0.05）。综上，混合油脂日粮相较于大豆油日粮提高了肉鸡的饲料转化效率，促进了胸肌组织中DHA和EPA的沉积；日粮油脂干预后肉鸡胸肌中磷脂代谢物、氧化稳定性代谢物和脂类代谢途径发生显著改变。本研究鉴定出的差异代谢物和差异代谢通路可为调控肉鸡脂肪酸组成的研究提供参考。     

L-Leucine In Ovo Administration Causes Growth Retardation and Modifies Specific Amino Acid Metabolism in Broiler Embryos

L-Leucine (L-Leu) in ovo administration was demonstrated to afford thermotolerance and modified amino acids metabolism in post-hatched broiler chicks under heat stress. This study aimed to investigate the changes in embryonic growth and amino acid metabolism after in ovo injection of L-Leu. Fertilized broiler eggs were subjected to in ovo injection of sterile water or L-Leu on embryonic day (ED) 7. The weight of embryos and yolk sacs were measured on ED 12, 14, 16, and 18. Plasma and livers were collected on ED 14 and 18 for free amino acid analysis. The weight and relative weight of embryos were significantly lowered by in ovo administration of L-Leu, but those of yolk sacs were not altered. Moreover, L-Leu in ovo injection significantly reduced the plasma proline concentration during embryogenesis and increased the plasma concentrations of tyrosine (Tyr) and lysine (Lys) in ED 18. Hepatic Lys concentration was also significantly increased by L-Leu in ovo injection. Interestingly, Leu concentrations in the plasma and liver were not affected by L-Leu administration. These results indicated that in ovo administered L-Leu was metabolized before ED 14 and affected embryonic growth and amino acid metabolism during embryogenesis.     

丁酸梭菌对高胆碱饮食小鼠血浆游离脂肪酸组成的影响

【目的】分析丁酸梭菌对高胆碱饮食造成脂代谢异常小鼠血浆游离脂肪酸组成(FFA)的影响,以阐明丁酸梭菌调节高胆碱膳食脂代谢的作用机制。【方法】选取4周龄健康的雄性昆明小鼠24只,随机分为3组：正常组、模型组和丁酸梭菌组,每组8只。高胆碱饮食造模8周后,给药7 d,禁食12 h,采集血浆、肝脏、附睾脂肪垫和肾周脂肪,对肝脏、脂肪称重,利用生化仪检测血脂水平;通过HE染色及油红O染色分别观察肝脏组织结构变化及脂滴沉积程度;利用液相色谱-质谱联用(LC-MS)技术检测小鼠血浆氧化三甲胺(TMAO)含量;利用气相色谱-质谱联用(GC-MS)技术及多元统计分析研究血浆中FFA组成。【结果】与模型组相比,丁酸梭菌组小鼠体重、脂肪增长得到显著抑制(P<0.05),给予丁酸梭菌后小鼠肝脏脂肪沉积减少;丁酸梭菌显著或极显著降低了高胆碱饮食小鼠血浆中甘油三酯(TG)和低密度脂蛋白(LDL-C)含量(P<0.05;P<0.01),并使高密度脂蛋白(HDL-C)含量显著升高(P<0.05);丁酸梭菌灌胃后,高胆碱饮食小鼠血浆TMAO浓度显著降低(P<0.05);丁酸梭菌可显著降低高...     

Regulation Effects and Mchanism of Amino Acids on Lipid Metabolism in Mammals

Lipid metabolism not only affects human health but also is vital to livestock and poultry production. Therefore, it is necessary to study the nutrition regulation strategy of lipid metabolism. This paper mainly introduces the four amino acids which are important for the regulation of lipid metabolism, including methionine (Met),leucine (Leu),arginine (Arg) and glutamate (Glu), and summarizes the regulation effects and mechanism on lipid metabolism of the four amino acids at the physiological level, cellular level and molecular level. This paper introduces these four amino acids in the role of nutritional research, physiological effects in animal experiments,the mechanism of regulation effect on lipid metabolism and the combination effect. Meanwhile,the regulation and mechanism of amino acid on lipid metabolism in adipose,liver and muscle tissue were emphatically discussed. This paper reviews the progress of four amino acids in basic research and livestock production application, and further analyze the potential directions in the future research on the deep mechanism of lipid metabolism to clarify the regulation effects and potential mechanism of the amino acids on mammals' lipid metabolism, which was aimed to provide a reference for the accuracy using of amino acids.     

氨基酸对哺乳动物脂质代谢的调控及其作用机理

脂质代谢不仅影响人类健康,对畜禽生产也至关重要,研究脂质代谢的营养调控策略有重要意义。作者主要介绍了蛋氨酸（Met）、亮氨酸（Leu）、精氨酸（Arg）和谷氨酸（Glu）这4种对脂质代谢调控作用显著的氨基酸。从生理水平、细胞水平和分子水平上总结了4种氨基酸对哺乳动物脂质代谢的调控及其作用机理,并从4种氨基酸在营养学研究中的作用、在动物试验中的生理效应、对脂质代谢调控的作用机理及各种氨基酸的组合效应等方面进行综述,重点综述了氨基酸对脂肪、肝脏和肌肉组织脂质代谢的调控及其作用机理。同时回顾了4种氨基酸在基础研究与畜牧生产应用上取得的进展,进一步分析了未来研究氨基酸对脂质代谢调控深层机理的可能方向,以期为氨基酸的精准利用提供参考。     

Effect of short chain fatty acids on the performance and intestinal weight in germ-free and conventional chicks

1. In experiment 1, the performance and tissue weights of germ-free (GF) and conventional (CV) chicks fed on diets containing 25.4 g acetic acid/kg diet (AD) or 25.4 g kaolin/kg diet (KD) were investigated. Body weight gain in GF chicks was significantly higher on the AD, but significantly lower on the KD compared with their CV counterparts. The values for food efficiency, protein retention and energy retention followed a similar pattern to that of the body weight gain. 2. The weights of all sections of the intestine except the colon were significantly greater in CV chicks. In CV but not in GF birds the jejunum and ileum were heavier from birds fed on the AD than from those on the KD diet. 3. In experiment 2, the influence of butyric acid administration on the weight of some organs in chicks was investigated. The weight of duodenum, jejunum and ileum was significantly increased by intraperitoneal administration of butyric acid (2 ml of 100 mM solution/d) for 4 d, but no significant effect was observed by oral administration. 4. It might be suggested that short chain fatty acids such as acetic and butyric acids formed by bacterial action in the crop and subsequently absorbed are at least partly responsible for the heavier gut weight in CV birds.     

Effects of acetylated wood powder on growth performance,hepatic and muscular free amino acid profiles,and inosine 5’-monophosphate concentration of breast meat in broiler chickens

1. The present study was conducted to determine the effects of acetylated wood powder (AW) as a new feed additive on performance, liver and muscle metabolism of amino acids and fatty acids and nucleotide-related substances of meat in broiler chickens. It was hypothesised that acetic acid desorbed from AW during intestinal digestion affects tissue metabolism.

2. Two-week-old broiler chicks were divided into four groups and fed on diets supplemented with wood powder (30 g/kg) less than 106 µm in diameter, except for controls. The AW was added to diets at 0, 10 and 30 g/kg to replace the non-acetylated wood powder (NAW) for 26 d. Plasma, liver tissue and breast muscle were taken from half of birds at 40 d of age under the fed condition. After the remaining chickens were fasted for 14 h, breast muscle was taken and refrigerated for 24 h.

3. Consumption of wood powder with or without acetyl groups had no effect on growth performance including tissue weights of abdominal fat and breast muscle and plasma metabolites.

4. Feeding AW decreased total free amino acid concentrations in the liver compared to the group only fed on the NAW. This response was dependent mainly on reduced non-essential and glucogenic amino acid concentrations. However, in breast muscle, alterations of free amino acid concentrations were observed only for histidine and tryptophan. In addition, the fatty acid composition of liver and breast muscle was not affected by feeding AW.

5. In breast meat obtained from fasted chickens, the higher level of AW increased the concentration of inosine 5′-monophosphate, a taste-active compound, and in contrast, decreased the subsequent catabolites (inosine and hypoxanthine). However, the concentration of glutamic acid, a taste-active compound, was lowered at this level of AW ingestion.

6. Therefore, this study suggested that feeding AW as a new feed additive regulates ante-mortem amino acid utilisation in the liver and contributes to retard post-mortem degradation of inosine 5′-monophosphate as a taste-active compound in chicken meat.

     

限时采食对猪生长性能和肝代谢的影响

旨在研究限时采食对猪生长性能和肝转录谱及代谢谱的影响。本试验选择体重相近((56.29±1.93) kg)、健康的杜×长×大三元杂交猪12头随机分为对照组和限时采食组。每组6个重复,每个重复1头猪。对照组(CON)自由采食;限时采食组(TRF)每天饲喂3次(分别在7:00、12:00、17:00),每次饲喂足量饲料,每次采食时间为1 h。试验共计21 d,每天记录采食量,每周称重,计算料重比。试验结束屠宰全部试验猪,采集静脉血测定血液生化指标,采集肝用于代谢组和转录组测定。结果显示,与对照组相比,限时采食组猪平均日增重显著升高(P<0.05),料重比显著降低(P<0.05)。与对照组相比,限时采食组猪血清总胆汁酸浓度有下降趋势(P<0.1),而丙氨酸转氨酶和肌酸激酶的浓度有上升趋势(P<0.1)。限时采食显著增加抑制蛋白质分解基因的表达(P<0.05),减少肝总蛋白质的分解,使肝中氨基酸水平下降。此外,限时采食还显著上调脂肪酸代谢相关基因的表达(P<0.05),增加肝脂肪的合成,减少肝脂肪酸含量。综上所述,限时采食可以影响肝中的氮代谢和氨基酸代谢,改变脂肪酸和蛋白质在肝中的代谢,进而调控生长猪对营养物质的利用并影响猪的生长性能,这提示可以通过饲喂模式改善动物生长和代谢。     

Regulation of free glutamate content in meat by dietary lysine in broilers

Regulation of taste is important for improving meat quality and glutamate (Glu) is one of the important taste‐active components in meat. Here, the effects of dietary lysine (Lys) content on taste‐active components in meat, especially free Glu, were investigated. Fourteen‐day‐old broiler chicks (Gallus gallus) were fed on diets containing 100% or 150% of the recommended Lys content for 10 days. Concentrations of free amino acids in plasma, muscle and liver were measured. The levels of messenger RNAs (mRNAs) for enzymes related to Glu metabolism were determined in muscle and liver. The concentration of muscle metabolites was also determined. The free Glu content in muscle of chicks fed the Lys150% diet was increased by 44.0% compared with that in chicks fed the Lys100% diet (P < 0.01). The mRNA level of lysine α‐ketoglutarate reductase, which is involved in Lys degradation and Glu production, was significantly increased (P < 0.05) in the Lys150% group. Metabolome analysis showed that the Lys degradation products, muscular saccharopine, pipecolic acid and α‐aminoadipic acid, were increased in the Lys150% group. Our results suggest that free Glu content in muscle is regulated by Lys degradation. These results suggest that a short‐term feeding of high‐Lys diet could improve the taste of meat.     

Metabolomic differences of seminal plasma between boars with high and low average conception rates after artificial insemination

Seminal plasma is a complex biological fluid containing many metabolites including amino acids, fructose, carbohydrates and lipids Metabolites play important roles in multiple biological processes, but details and significance of the seminal plasma metabolome related to boar fertility are unknown. The aim of the present study was to compare the comprehensive metabolome of seminal plasma from boars with different conception rate after artificial insemination and to identify the potential biomarkers. Semen samples were collected from boars which divided into two groups according to the conception rates in the offspring. Seminal plasma metabolites were isolated, purified, and then subjected to Ultra-high Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-qTOF-MS) procession. A total of 576 (Positive ion mode) and 377 (Negative ion mode) metabolites were identified in seminal plasma. Metabolites were identified and categorized according to their major chemical classes, including carboxylic acids and derivatives, organooxygen compounds, amino acids, peptides, and alogues, fatty amides, fatty acyls, benzene and substituted derivatives, purine nucleotides, pyrimidine nucleotides, glycosyl compounds, fatty acids and conjugates. The results showed that 4-Aminobenzoate, Pro-Asn, Ile-Tyr, Homoveratric acid and D-Biotin were higher in semen of boar with higher conception rate (HG) versus lower conception rate (LG) (p < .05), whereas L-Serine, Butoxyacetic acid, S-Methyl-5'-thioadenosine, Capsaicin and 1-O-(cis-9-Octadecenyl)-2-O-acetyl-sn-glycero-3-phosphocholine (PAF) were lower in HG than in LG (p < .05). These metabolites may be considered as candidate biomarkers for different fertility in boars.     

内蒙古白绒山羊的限制性氨基酸研究

本文旨在对内蒙古白绒山羊限制性氨基酸进行研究。选用12只装有瘤胃、十二指肠和回肠瘘管的内蒙古白绒山羊,采用随机区组试验设计,并采用递减法进行十二指肠氨基酸灌注,试验分别测定了小肠可吸收氨基酸的量、氨基酸的平衡性、氮沉积、日增重等指标。试验结果表明:(1)减少蛋氨酸、组氨酸、精氨酸、丝氨酸、半胱氨酸对内蒙古白绒山羊的小肠氨基酸消失量、氨基酸的平衡性均有显著影响。(2)氨基酸灌注量减少后,根据氮沉积求出的各种氨基酸的相对限制程度依次为半胱氨酸(72.17%)、丝氨酸(54.25%)、精氨酸(51.76%)、蛋氨酸(29.07%)、组氨酸(20.45%)。内蒙古白绒山羊的限制性氨基酸是半胱氨酸、丝氨酸、精氨酸、蛋氨酸和组氨酸。     

Essential Fatty Acids and the Cat

Abstract— The physiology of essential fatty acid metabolism in the cat is reviewed. Emphasis is placed on those aspects of the n:6 and n:3 fatty acids, their metabolites and interactions, which relate primarily to the skin. The functional roles, if known, of the fatty acids are discussed. Recent clinical research into the use of essential fatty acid supplements in the management of feline dermatoses is presented. Current indications for the therapeutic supplementation with essential fatty acids are summarised.     

Effect of different time intervals after feeding on plasma metabolites in growing pigs: an UPLC‐MS‐based metabolomics study

A diet consumed by pigs provides the nutrients for the production of a large number of metabolites that, after first‐pass metabolism in the liver, circulate systemically where they may exert diverse physiologic influences on pigs. So far, little is known of how feeding elicits changes in metabolic profiles for growing pigs. This study investigated differences in plasma metabolites in growing pigs at several intervals after feeding using the technique of metabolomics. Ten barrows (22.5 ± 0.5 kg BW) were fed a corn‐soybean meal basal diet and were kept in metabolism crates for a period of 11 days. An indwelling catheter was inserted into the jugular vein of each pig before the experimental period. Plasmas before and 1, 4, and 8 hr after feeding were collected at day 11 and differential metabolites were determined using a metabolomics approach. Direct comparison at several intervals after feeding revealed differences in 14 compounds. Identified signatures were enriched in metabolic pathways related to linoleic acid metabolism, arginine and proline metabolism, lysine degradation, glycine, serine and threonine metabolism, and lysine biosynthesis. These results suggest that plasma metabolites of growing pigs after feeding were modulated through changes in linoleic acid metabolism and amino acid metabolism.     

亚急性瘤胃酸中毒耐受性不同的奶牛血浆和乳中脂肪酸及代谢物组成分析

为比较研究对亚急性瘤胃酸中毒（SARA）耐受性不同的奶牛血浆和乳中代谢组的差异，试验选用12头装有永久性瘤胃瘘管的泌乳中期荷斯坦奶牛［泌乳天数=（114±22） d］，饲喂精粗比为4∶6的全混合日粮。在试验期第20和21天晨饲后0、2、4、6、8和12 h监测瘤胃pH值，在早晚饲喂前收集奶样，晨饲6 h后采集颈静脉血，用于测定脂肪酸及代谢物组成。基于奶牛瘤胃平均pH值高低，将奶牛分为SARA易感组（SUS，瘤胃pH=5.76，n=4）和SARA耐受组（TOL，瘤胃pH=6.10，n=4）。脂肪酸测定结果分析显示，与TOL组相比，SUS组奶牛血液及乳中的碳链长度≤C16的脂肪酸含量显著增高（P<0.05），而>C16的脂肪酸的含量显著降低（P<0.05），乳中饱和脂肪酸及单不饱和脂肪酸含量均显著降低（P<0.05）。血浆代谢组结果显示， SUS组奶牛血浆中L-苯丙酮酸水平显著降低（P<0.05），而MG（18：0/0：0/0：0）、9-HODE、12（13） Ep-9-KODE、烟酰胺、异戊基肉碱、磷酸肌酸和L-谷氨酸水平显著升高（P<0.05）。乳代谢组分析结果显示，与TOL组比较，SUS组奶牛乳中的1-硬脂酰磷酸甘油丝氨酸和鞘氨醇水平显著升高（P<0.05），而甘油磷酰基乙醇胺、3-磷酸甘油和乳清酸等其他14个差异代谢物水平显著降低（P<0.05），这些差异代谢物主要涉及甘油磷脂代谢、甘油脂代谢及泛酸与辅酶A代谢等代谢途径。综上所述，SARA耐受性不同的奶牛的血浆和乳中脂肪酸及代谢物组成存在较大差异，SUS组奶牛血浆中氨基酸代谢活动增加，乳腺的乳脂从头合成能力较强，而从血液中摄取长链脂肪酸能力减弱，与此同时，易感奶牛乳中磷脂、乳清酸等营养物质水平下降，乳品质下降。     

The effect of proteinase inhibitors in rats. 1. Report. Effect of a short-term application on the N balance, the free amino acids in liver and muscles and the biological half-life of the intestinal tissue proteins]

Short-term trials with a triazin derivative and a peptide aldehyde were carried out to investigate the in-vivo administration of proteinase inhibitors in rats for improving the N balance of the animals. The results of N balances suggest that the peptide aldehyde may be regarded as a promising substance that may be used to increase the rate of utilization of the absorbed N in the intermediary metabolism. The short-term use of both active compounds did not affect the true N digestibility. The two compounds did not influence the concentration of free amino acids in the liver and muscle tissue under the experimental feeding conditions used. Similarly, tests of the biological half-life of proteins taken from the small and large intestine did not reveal any changes due to the presence of the inhibitors. Animals receiving a protein-free diet exhibited longer half-life periods for proteins from the small and large intestine compared with animals of the other groups. A protein-free nutrition of the animals caused a general increase in the levels of free non-essential amino acids in the liver. This was accompanied by a simultaneous decrease in essential amino acids. The same type of nutrition led to a general decline in the concentration of free amino acids in muscular tissue, particularly of the essential amino acids. High-protein nutrition led especially to an increase in the concentration of branched-chain amino acids in both types of tissues investigated.